Treatment of metastatic breast cancer

ABSTRACT

The present invention relates to the use of an anti-EpCAM antibody for the manufacture of a medicament for the treatment of metastatic breast cancer. The present invention further relates to a method of treating metastatic breast cancer comprising administering said anti-EpCAM antibody.

This application is a continuation of U.S. application Ser. No.13/180,093 filed Jul. 11, 2011, which is a continuation of U.S.application Ser. No. 12/162,102 filed Dec. 19, 2008 now issued U.S. Pat.No. 7,976,842 on Jul. 12, 2011, which is a national phase applicationunder 35 U.S.C. §371 of International Application No. PCT/EP2007/001127filed Feb. 9, 2007, which claims priority to European Patent ApplicationNo. EP 06 002 680.4 filed Feb. 9, 2006 and U.S. Provisional ApplicationNo. 60/772,421 filed Feb. 9, 2006. The entire text of each of theabove-referenced disclosures is specifically incorporated herein byreference without disclaimer.

REFERENCE TO THE SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing is provided as a filed entitled“47306B SubSeqListing.txt” created Mar. 8, 2013 which is 8 KB in size.The information in the electronic format of the Sequence Listing isincorporated herein by reference in its entirety.

The present invention relates to the use of an anti-EpCAM antibody forthe manufacture of a medicament for the treatment of metastatic breastcancer. The present invention further relates to a method of treatingmetastatic breast cancer comprising administering said anti-EpCAMantibody.

Breast cancer is the most common cancer and the second cause of cancerdeath in women. In 2001, the incidence rates of breast cancer were90-100/100,000 in the United States and 50-70/100,000 in Europe. Theincidence of the disease is growing worldwide. Risk factors for breastcancer include race, age, and mutations in the tumor suppressor genesBRCA-1 and -2 and p53. Alcohol consumption, fat-rich diet, lack ofexercise, exogenous post-menopausal hormones and ionizing radiation alsoincrease the risk of developing breast cancer. Estrogen receptor andprogesterone receptor negative breast cancer (“ER-” and “PR-”,respectively), large tumor size, high grade cytology and age below 35years are associated with a bad prognosis (Goldhirsch et al. (2001). J.Clin. Oncol. 19: 3817-27). In 2005 an estimated 212,000 new cases ofinvasive and 58,000 new cases of non-invasive breast cancer will bediagnosed, with 40,000 women expected to die from breast cancer.

Breast cancer may generally be divided into several main stages: early,locally advanced, locally recurrent and metastatic. Early breast cancerincludes non-invasive breast cancer, for example lobular carcinoma insitu (“LCIS”) and ductile carcinoma in situ (“DCIS”). Most commonly,breast cancer is staged according to the Tumor Node Metastasis (“TNM”)classification system proposed by the American Joint Committee on Cancer(AJCC Cancer Staging Manual, 6^(th) Edition). The TNM classificationsystem defines 7 separate stages of breast cancer: 0, I, IIA, IIB, IIIA,IIIB and IV. Stages 0, I and subtypes of stage II are generallyassociated with early stage breast cancer. Stage III as well as subtypesof stage II are generally associated with advanced breast cancer. StageIV is generally associated with metastatic breast cancer. More detailedinformation on the TNM classification of breast cancer is shown inFIG. 1. Tumor size may be measured and monitored by the ResponseEvaluation Criteria in Solid Tumors (“RECIST”) criteria (Therasse et al.(2000). J. Natl. Cancer Inst. 92: 205-16).

While the 5-year survival prognosis for early stage breast cancer isgenerally above 60%, this number drops to between 40-60% for advancedbreast cancer. The 5-year survival prognosis is generally around 15% formetastatic breast cancer. The most common sites of distant metastasisfor breast cancer include lung, liver, bone, lymph nodes, skin and CNS(brain). Once metastatic breast cancer has been diagnosed, a patient mayon average expect to live a further 18-24 months. Cure for metastaticbreast cancer is unlikely, and the modes of therapy for this systemicdisease are largely palliative in nature.

The above emphasizes the importance of new developments in breast cancertherapy, especially therapy for metastatic breast cancer.

Current therapeutic options for treatment of breast cancer, includingmetastatic breast cancer, include surgery (e.g. resection, autologousbone marrow transplantation), radiation therapy, chemotherapy (e.g.anthracyclines such as doxorubicin, alkylating agents such ascyclophosphamide and mitomycin C, taxanes such as paclitaxel anddocetaxel, antimetabolites such as capecitabine, microtubule inhibitorssuch as the vinca alkaloid navelbine), endocrine therapy (e.g.antiestrogens such as tamoxifen, progestins such as medroxyprogesteroneacetate and megastrol acetate, aromatase inhibitors such asaminoglutethamide and letrozole) and biologics (e.g. cytokines,immunotherapeutics such as monoclonal antibodies). Most commonlymetastatic breast cancer is treated by one or a combination ofchemotherapy (the most effective drugs including cyclophosphamide,doxorubicin, navelbine, capecitabine and mitomycin C) and endocrinetherapy.

The standard care in breast cancer is surgical removal of the tumor andradiotherapy, preceded or followed by either hormonal therapy orchemotherapy, depending on the tumor stage and risk factors. Patientswith stage I to stage IIIA (see below, as well as FIG. 1) may be treatedwith adjuvant chemotherapy or hormone therapy. In patients withinoperable invasive stage IIIB disease or stage IV metastatic breastcancer, chemotherapy merely alleviates the symptoms.

Recently taxanes and anthracyclines have markedly improved the survivalrates of breast cancer patients. Capecitabine (Xeloda®, capecitabine,Roche Ltd: Summary of Product Characteristics) has been approved forsecond-line or higher treatment in patients who have failed cytotoxicchemotherapy including anthracyclines and/or taxanes, especially becauseof its low toxicity and oral formulation (O'Shaughnessy (2002). Oncology16: 17-22). However, despite these improved treatment modalities,survival of patients with advanced breast cancer remains poor andchemotherapy is only palliative.

The monoclonal anti-Her-2/neu antibody trastuzumab (Herceptin®,trastuzumab, Roche, Ltd: Summary of Product Characteristics, March 2002)was the first biological targeted therapy approved for treatment ofpatients with breast cancer whose tumors overexpress Her-2/neu. Incombination with paclitaxel, it is indicated as a first-line treatmentof patients with metastatic breast cancer and as second-line or highertreatment as single agent in the same patient population (Cardoso et al.(2002). Clin. Breast Cancer 3: 258-9; Tan-Chiu & Piccart (2002).Oncology 63: 57-63). However, only a small fraction of patients withbreast cancer (approximately 20%) overexpress Her2/neu at a high level,and are therefore eligible for treatment with this antibody.

Therefore, development of new anti-cancer drugs, especially for thetreatment of breast cancer patients in whom trastuzumab is notindicated, is an important medical need.

One promising immunotherapeutic is the human antibody comprising in itsheavy chain variable region amino acid sequences as set out in SEQ IDNOs. 3, 4 and 5, and/or comprising in its light chain variable regionamino acid sequences as set out in SEQ ID NOs. 6, 7 and 8. Hereinafterthis antibody will be referred to as “Anti-EpCAM”, and is furthercharacterized in its heavy and light chains by the amino acid sequencesas set out in SEQ ID NOs. 1 and 2, respectively). This antibody binds tothe epithelial cell adhesion molecule (“EpCAM”, also called 17-1Aantigen, KSA, EGP40, GA733-2, ks 1-4 and esa). EpCAM is a highlyconserved surface glycoprotein which is overexpressed in many carcinomasof different origins, including breast cancer. Tumor samples from 3722patients with colon, stomach, lung, ovarian or prostate cancer wereanalyzed for EpCAM expression using a sensitive immunohistochemicalstaining assay on tissue microarrays. An intermediate to strong EpCAMexpression was reported in more than 88% of all tumor samples, in 94% ofovarian cancers, 94% of colon cancers, 92% of stomach cancers, 90% ofprostate cancers and in 71% of lung cancers. These results confirm thatEpCAM is frequently present in epithelial tumor cells, and highlightsAnti-EpCAM as a potential diagnostic and therapeutic target.

In two studies in primary breast cancer tumors, strong EpCAM expressionwas shown in 36% of 384 sections (Tandon et al. (1990) Cancer Res. 50:3317-24) and 59% of 128 samples (Edwards et al. (1986) Cancer Res. 46:1306-17), respectively. In another study (Spizzo et al. (2002) Int. J.Cancer 98: 883-8), strong EpCAM expression was found in 73 of 205 (36%)primary breast tumor specimens and the authors report that EpCAMoverexpression in breast cancer was associated with reduced disease-freeand overall survival. EpCAM overexpression was also correlated withtumor size and with hormone receptor negativity; it was highest inductal breast cancers as well as in histological grade III subtypes. Inanother series, approximately 90% of breast cancer samples were shown toexpress EpCAM to some extent and more than 40% showed strong EpCAMexpression.

In vitro, Anti-EpCAM causes both antibody-dependent cellularcytotoxicity (“ADCC”) and complement-dependent cytotoxicity (“CDC”). Asthe most likely mechanism of action, Anti-EpCAM recruits the patient'snatural killer cells to the tumor site by binding to EpCAM-positivetumor cells. Via activation of the patient's immune effector cells,EpCAM-positive tumor cells can then be eliminated.

Certain therapeutic regimens employing Anti-EpCAM are known in the art(WO 2005/080428). Specifically, WO 2005/080428 describes a therapeuticregimen involving administration of Anti-EpCAM to cancer patients. Here,it was contemplated to administer Anti-EpCAM in the treatment of, forexample, breast cancer or a minimal residual disease. In the context ofthe latter, a minimal residual disease may be understood as the localand non-local reoccurrence of a tumor caused by the survival of singletumor cells.

It is a goal of the present invention to improve upon existing breastcancer therapies.

Accordingly, one aspect of the invention relates to a use of ananti-EpCAM antibody comprising amino acid sequences as set out in SEQ IDNOs. 3, 4, 5, 6, 7 and/or 8 (“Anti-EpCAM”), for the manufacture of amedicament for the treatment of human metastatic breast cancer.

As used herein, the term “metastatic breast cancer” is to be understoodas a disease in which at least one transformed, i.e. cancerous cell froma primary tumor of the breast has become separated from the primarytumor and has continued to grow into a tumor at a location distinct fromthat of the primary tumor (hereinafter “distinct location”). Thedistinct location may for example be within the same breast as that inwhich the primary tumor is located (ipsilateral breast) or within theother breast (contralateral breast). As further examples, the distinctlocation may be within one or more lymph nodes, whether these aremovable or fixed, ipsilateral or contralateral to the primary tumor,supraclavicular, axillary or otherwise. Within the context of the TNMtumor classification system (shown in FIG. 1), a “metastatic breastcancer” as used herein would include i.a. all tumors whose stagingincludes M=1 (i.e. Stage 1V breast cancer; see FIG. 1), i.e. all tumorsfor which any degree of metastasis exists to distant locations such asfor example lung, liver, bone, lymph nodes, skin, brain and/or adistinct location within an ipsilateral and/or contralateral breast.

The term “breast cancer” as used herein denotes a disease in which aprimary tumor or multiple individual primary tumors exist in the breastor breasts. Generally, this means that no cancerous cell has (yet)become separated from the primary tumor in the breast, and has notspread to a “distinct location”. In this context, it should be notedthat the existence of multiple primary tumors within the same or withinboth the ipsilateral and contralateral breasts is not in itself to beunderstood as falling within the meaning of “metastatic breast cancer”.This is because multiple cells within one or both breasts may give riseto multiple primary tumors, none of which are or have yet becomemetastatic. On the other hand, the separation of a cancerous cell fromonly one of multiple primary tumors within a single breast and thesubsequent development of this single cell at a “distinct location” intoa separate tumor would constitute “metastatic breast cancer” as usedherein, irrespective of the presence or absence of one or more primarytumors in at least one of the breasts.

It should be noted that the term “metastatic breast cancer” as usedherein does not imply that said metastasis existing at a “distinctlocation” must have arisen from any one particular primary tumor of thebreast. That is to say, the origin of the metastasis at the “distinctlocation” is immaterial to the designation of the disease as “metastaticbreast cancer” as long as the primary tumor giving rise to themetastasis originated in the breast tissue. For this purpose, the term“breast tissue” is to be understood as including the lobules and theducts of the breast, i.e. the tissue which most commonly gives rise totumors of the breast.

The applicant has surprisingly found that Anti-EpCAM is well suited notonly to the treatment of breast cancer as such, i.e. breast cancerinvolving at least one primary tumor in the breast, but also to thetreatment of metastatic breast cancer. That Anti-EpCAM can be used inthis fashion is not at all to be expected, as the tumor load (i.e. thenumber of cancer cells, the size of a tumor, or the amount of cancer inthe body; also called “tumor burden”) associated with metastatic breastcancer is generally greater than that observed in non-metastatic breastcancer. This is because a single primary breast tumor may well—and oftendoes—engender multiple dispersed metastases throughout the body. Theabsolute number of EpCAM molecules existing on the surface of malignantcells throughout the body is therefore generally greater in metastaticbreast cancer than in non-metastatic breast cancer. The data provided inthe appended examples show that the administration of a pharmaceuticalcomposition comprising an anti-EpCAM antibody leads to a significantprolongation of the TTP (time to progression) of the treated disease.The actual effect appears to be correlated with the expression level ofEpCAM on the surface of the malignant cells to be treated. Patients maybe sorted into groups of different groups of EpCAM expressors accordingto Gastl et al. (2000) Lancet 356, 1981-2. Briefly, Gastl et al.analyzed the EpCAM expression of tumor cells isolated from differentpatients in immunohistochemical stains. A total immunostaining score wascalculated as the product of a proportion score and an intensity score.The proportion score describes according to Gastl et al. the estimatedfraction of positive-stained tumor cells (0, none; 1, <10%; <10%; 2,10%-50%; 3, 50%-80%; 4, >80%). The intensity score represented accordingto Gastl et al. the estimated staining intensity (0, no staining; 1,weak; 2, moderate; 3, strong). The resulting total score ranges from 0to 12. High EpCAM is defined as a total score greater than 4, since thepatient sample showed a bimodal distribution of EpCAM expression (lowand high EpCAM expressors) with the discriminating nadir at a totalscore value of 3 to 4. The prognosis for a patient receiving a therapycomprising the administration of an anti-EpCAM antibody, which isidentified as a high EpCAM expressor is more optimistic than for apatient identified as a moderate or low EpCAM expressor. In line withthis observation patients identified as extremely high EpCAM expressors(a patient showing a higher amount of EpCAM on the surface of malignantcells than the mean of high EpCAM expressors, i.e. a total score of ≧8)show a further prolongation of the TTP compared with the patientsidentified as high EpCAM expressors and receiving the same therapy.Moreover, the amount of anti-EpCAM antibody in a pharmaceuticalcomposition administered to a patient directly correlates with theprognosis. In particular, the administration of high dose of anti-EpCAMantibody results in a prolongation of the TTP of the treated diseasecompared to the administration of a low dose to a patient of the samegroup of EpCAM expressors.

In one preferred embodiment the anti-EpCAM antibody Anti-EpCAM is ahuman antibody.

In a further preferred embodiment Anti-EpCAM comprises all of SEQ IDNOs. 3, 4, 5, 6, 7 and 8. In a further preferred embodiment, Anti-EpCAMcomprises SEQ ID NOs. 1 and/or 2. In an especially preferred embodiment,Anti-EpCAM comprises both SEQ ID NOs. 1 and 2.

According to a preferred embodiment, the treatment of metastatic breastcancer comprises long-term stabilization of metastatic breast cancer.“Long-term stabilization” is to be understood as the case in whichdisease progression is stabilized at or below its beginning level overthe course of treatment with the anti-EpCAM antibody. This may beunderstood as a prolongation of the time taken to disease progression.“Long-term stabilization” also comprises the scenario in which the tumorshrinks (partial response). “Long-term stabilization” also comprises thescenario in which disease progression is reduced to or below thedetectable level, i.e. the patient responds completely to treatment andthe disease is cured (complete response). In such a scenario, treatmentmay be continued indefinitely as needed to prevent recurrence of thedisease, or may be terminated at the physician's discretion.

According to a preferred embodiment, the medicament is suitable for aso-called “low dose administration”. For the low dose administration thedose of each administration is in a range of 1 to 3 mg anti-EpCAMantibody/kg body weight. Preferably, the low dose administrationcomprises at least one loading dose in a range of 1 to 3 mg/kg bodyweight, followed by multiple maintenance doses, each maintenance dosebeing in a range of 1 to 3 mg/kg body weight. It is also preferred thatthe individual doses for the low dose administration are in a range of1.5 to 2.5 mg/kg body weight, more preferably, in a range of 1.75 to2.25 mg/kg body weight. Most preferably, the individual doses for thelow dose administration is 2 mg/kg body weight. It is preferred for thelow dose administration that the at least one loading dose for the lowdose administration is of 2 mg/kg body weight followed by multiplemaintenance doses, each maintenance dose being 2 mg/kg body weight.Alternatively, the medicament is suitable for a so-called “high doseadministration”. For the high dose administration the dose of eachadministration is in a range of 4.5 to 8 mg anti-EpCAM antibody/kg bodyweight. Preferably, the high dose administration comprises at least oneloading dose in a range of 4.5 to 8 mg/kg body weight, followed bymultiple maintenance doses, each maintenance dose being in a range of4.5 to 8 mg/kg body weight. It is also preferred that the individualdoses for the high dose administration are in a range of 5 to 7 mg/kgbody weight, more preferably, in a range of 5.5 to 6.5 mg/kg bodyweight, further preferably, in a range of 5.75 to 6.25 mg/kg bodyweight. Most preferably, the individual doses for the high doseadministration is 6 mg/kg body weight. It is preferred for the high doseadministration that the at least one loading dose for the high doseadministration is of 6 mg/kg body weight followed by multiplemaintenance doses, each maintenance dose being 6 mg/kg body weight. Itis unexpected that such loading and maintenance doses would yield atherapeutic benefit for the treatment of metastatic breast cancer, inwhich multiple metastases throughout the body are to be eradicated.

According to a further embodiment of the invention, it may beadvantageous to establish whether a metastatic breast cancer patientexpresses EpCAM to a higher or to a lower extent. Patients may be sortedinto groups of non-EpCAM expressors, moderate EpCAM expressors, lowEpCAM expressors and high EpCAM expressors as for example described inGastl et al. (2000) Lancet 356, 1981-2. In general, it can beadvantageous to correlate the amount of Anti-EpCAM administered to ametastatic breast cancer patient with the level of EpCAM expressionobserved for the patient in question, with high EpCAM expressorsreceiving higher doses of Anti-EpCAM and low EpCAM expressors receivinglower doses of Anti-EpCAM. It can be especially advantageous toadminister to patients expressing high levels of EpCAM the higher of thetwo above doses of Anti-EpCAM, namely 6 mg/kg body weight.

According to a further embodiment of the invention, the duration of timebetween a respective loading dose and either another successive loadingdose or a first maintenance dose is to be no longer than a week, whilethe duration of time between a respective maintenance dose and afollowing maintenance dose is to be no longer than two weeks.Preferably, the/each loading dose/s is/are administered every week andeach of said maintenance doses is administered every second week. Weeklyadministration of loading doses of Anti-EpCAM in the “loading phase”ensures that the minimum level of Anti-EpCAM in serum (taking account ofcontinual clearance in the form of excretion and elimination) remainshigh enough at all times to elicit the desired therapeutic effect. Thisminimum level of Anti-EpCAM required for therapeutic effect is known asthe “serum trough level”, and will be referred to as such hereinbelow.Once this serum level is reached, further administration of maintenancedoses of Anti-EpCAM in the subsequent “maintenance phase” in two-weekintervals ensures (again, taking account of continual clearance) thatthe serum level of Anti-EpCAM never sinks below what is required for acontinued therapeutic effect. The pharmacokinetic calculations requiredto determine the serum trough level for Anti-EpCAM are described in theart (see WO 2005/080428).

According to an especially preferred embodiment, one loading dose isadministered at the beginning of each of therapy weeks 1, 2 and 3followed by 11 maintenance doses, one maintenance dose beingadministered at the beginning of each of therapy weeks 4, 6, 8, 10, 12,14, 16, 18, 20, 22 and 24. Here, it has been surprisingly found that thecombination of 3 loading doses in the above time intervals followed by11 maintenance doses in the above intervals is especially effective intreating metastatic breast cancer. This implies a total therapeuticperiod of 24 weeks from start to finish (not taking into account anypost-treatment checkups normally associated with any kind of suchtherapies). In a further preferred embodiment the total therapeuticperiod from start to finish is of 30 weeks, 40 weeks, 50 weeks or 60weeks. It is also preferred that a period of administration of EpCAMantibody in line with the above described scheme is followed by a periodwithout administration of EpCAM antibody and a further therapeuticperiod of administration of EpCAM antibody. Such an order of periods maybe repeated for several times.

Alternatively, a further embodiment of the invention contemplatesadministration of Anti-EpCAM in loading doses as described above,followed by as many maintenance doses as required to control tumorprogression. Within this embodiment, tumor progression may be seen asbeing controlled as long as the size of one or more monitored metastatictumors is not increasing. In the best case, the size of one or moretumors monitored may actually shrink (as in a partial response). Here,the tumor(s) monitored may shrink to nothing, i.e. may disappear (as ina complete response). The tumor(s) monitored may remain the same sizeand the time to disease progression may thus be increased (as instabilized disease). According to this embodiment, therefore,maintenance doses of Anti-EpCAM may be continued in the above intervalsindefinitely as long as there is either a partial response or a stableresponse, continuing to the case in which a complete response ismeasured. In the case of further tumor progression (i.e. the size ornumber of the monitored tumor(s) increases during treatment), thetreatment with Anti-EpCAM may be terminated and, if appropriate, bereplaced by an alternate form of therapy.

According to a further embodiment of the invention, Anti-EpCAM isadministered as a solution comprising 0.9% sodium chloride.

According to a further embodiment of the invention, Anti-EpCAM isadministered to a metastatic breast cancer patient intravenously.

A further aspect of the invention is the use of Anti-EpCAM for thetreatment of metastatic breast cancer.

A further aspect of the invention relates to a method of treating humanmetastatic breast cancer, said method comprising administering to ahuman an anti-EpCAM antibody comprising SEQ ID NOs. 3, 4, 5, 6, 7 and/or8. This antibody is further characterized by heavy and light chainsamino acid sequences as set out in SEQ ID NOs. 1 and 2, respectively.

Preferred embodiments of the present method of treating metastaticbreast cancer are as set out above in the context of the inventive use;these embodiments apply to the present inventive method mutatis mutandi.

The invention will now be illustrated by the following figures andnon-limiting examples.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 Overview of the TNM Classification/Staging System for breastcancer

FIG. 2 Time To Progression (“TTP”) plot showing the probability of beingfree of disease progression vs. time for low and high administered dosesof Anti-EpCAM to 32 and 35 patients, respectively

FIG. 3 (“TTP”) plot showing the probability of being free of diseaseprogression vs. time for low and high administered doses of Anti-EpCAMto 27 and 40 patients, respectively

FIG. 4 (“TTP”) plot showing the probability of being free of diseaseprogression vs. time for high and low doses of Anti-EpCAM administeredto patients expressing both high and low levels of EpCAM

FIG. 5 (“TTP”) plot showing the probability of being free of diseaseprogression vs. time for high doses of Anti-EpCAM administered topatients expressing high levels of EpCAM, as compared to all otherpatients.

FIG. 6 (“TTP”) plot showing the probability of being free of diseaseprogression vs. time for low and high administered doses of Anti-EpCAMto 54 and 55 patients, respectively

FIG. 7 (“TTP”) plot showing the probability of being free of diseaseprogression vs. time for high and low doses of Anti-EpCAM administeredto patients expressing both high and low levels of EpCAM

FIG. 8 (“TTP”) plot showing the probability of being free of diseaseprogression vs. time for high doses of Anti-EpCAM administered topatients expressing high levels of EpCAM, as compared to low doses ofAnti-EpCAM administered to patients expressing low levels of EpCAM.

EXAMPLES

General

The following examples are intended to illustrate various aspects of theinvention and are in no way limiting to the invention's scope.Generally, the examples describe a clinical study program designed forthe fully human IgG1 antibody termed “Anti-EpCAM” as well as resultsfrom this clinical study program. The amino acid sequences of the first,second and third complementarity determining regions (CDRs) of the heavychain variable region of Anti-EpCAM are as set out in SEQ ID NOs. 3, 4and 5, respectively. The amino acid sequences of the first, second andthird CDRs of the light chain variable region of Anti-EpCAM are as setout in SEQ ID NOs. 6, 7 and 8, respectively. The amino acid sequence ofthe heavy chain of Anti-EpCAM is as set out in SEQ ID NO. 1 and theamino acid sequence of the light chain of Anti-EpCAM is as set out inSEQ ID NO. 2. Throughout the following examples, the following terms andabbreviations are used:

-   ADCC antibody-dependent cell-mediated cytotoxicity-   AE adverse event-   ALT alanine aminotransferase-   ANCOVA analysis of covariance-   AP alkaline phosphatase-   AST aspartate aminotransferase-   AUC area-under-the-curve-   BRCA breast cancer tumor suppressor gene-   CBA cytometric bead array-   CDC complement-dependent cytotoxicity-   CHO Chinese hamster ovary-   C_(min) minimum drug concentration-   CNS central nervous system-   CR complete Response-   CRF case report form-   CRP C-reactive protein-   CT computerized tomography-   CTCAE common terminology criteria for adverse events-   ECOG Eastern Cooperative Oncology Group-   ELISA enzyme-linked immunosorbent assay-   EpCAM epithelial cell adhesion molecule-   FAS full analysis set-   GCP good clinical practice-   GGT gamma-glutamyltransferase-   HAHA human anti-human antibodies-   HBsAg hepatitis B surface antigen-   HCV hepatitis C virus-   HIV human immunodeficiency virus-   ICF informed consent form-   IEC independent ethics committee-   INR international normalized ratio-   IRB institutional review board-   LDH lactic dehydrogenase-   NK natural killer-   OTR overall tumor response-   PK pharmacokinetic-   PP per protocol analysis set-   PR partial response-   PT prothrombin time-   PTT partial thromboplastin time-   PVP polyvinylpyrrolidone-   RBC red blood cell-   RECIST Response Evaluation Criteria in Solid Tumors-   SAE serious adverse event-   SAF safety analysis set-   SAP statistical analysis plan-   SD stable disease-   SGOT serum glutamic oxaloacetic transaminase-   SGPT serum glutamic pyruvic transaminase-   ULN upper limits of normal-   WBC white blood cell-   WHO World Health Organization

Example 1 Phase II Clinical Study Program Designed for Anti-EpCAMExample 1.1 Summary of Clinical Study

The clinical study designed for Anti-EpCAM is summarized in thefollowing table (Table 1).

TABLE 1 Clinical Study Summary Study Anti-EpCAM is a novel, fully humanIgG1 Rationale monoclonal antibody, derived from the repertoire of humanIgD-positive B cells, binding specifically to the epithelial celladhesion molecule (EpCAM). EpCAM is a highly conserved surfaceglycoprotein, which is overexpressed in many carcinomas of differentorigin, including breast cancer. Anti- EpCAM has been shown toeffectively kill breast tumor cells by antibody-dependent cellularcytotoxicity (ADCC) in preclinical experiments and to be safe in humans.This study will investigate whether Anti- EpCAM has anti-tumor activityin patients with metastatic breast cancer and if it could offer a noveltreatment option for these patients. Study A randomized, open-label,multicenter, parallel Design group, phase II study. The study isdesigned to evaluate the efficacy and safety of Anti-EpCAM over 24 weeksof therapy at two different doses with positive EpCAM testing. Thecentral randomization process will be stratified according to the EpCAMtest results performed at screening. Upon registration in one of theEpCAM strata, patients will be randomly assigned to either the low dosetreatment group or the high dose treatment group. ConcomitantProhibited: Medication Any concomitant anti-tumor therapy other than theinvestigational product Anti-EpCAM such as hormonal therapy, biologicaltherapy, chemotherapy, radiation therapy. Therapy with chronic systemichigh-dose corticosteroids and other immunosuppressive drugs. Any otherinvestigational agent. Duration of For each patient, the study consistsof a 4-week Patient screening period, a 24-week treatment period andParticipation/ a 4-week safety follow-up period and a final PK/ Durationof PD visit (12 weeks after end of therapy). the Study The estimatedaccrual period is 9 months and the total duration of the study isexpected to be 22 months. Measurements Each patient will have a maximumof 18 visits, and Evaluation including one screening visit, 14 visitsduring the treatment period, and one final PK/PD Assessment 12 weeksafter the end of therapy. Patients may also undergo unscheduled visitsin case of changes in their medical condition. Efficacy Efficacyevaluations will occur every 6 weeks after the first administration ofAnti-EpCAM until week 24 and every 8 weeks thereafter (during follow-upstudy), and will include the following: Thoracic CT scan or chest X-rayAbdominal CT scan or MRI Bone scintigraphy (for patients with bonemetastasis at screening) Responses must be confirmed at a follow-upevaluation not earlier than 4 weeks later. Safety A physical examinationor a symptom-directed examination, including vital signs, and laboratoryparameters (hematology, clinical chemistry, coagulation profile, andurinalysis) will be conducted at every visit, except visit 18. ECG willbe performed at screening. ECOG performance status will be assessed atscreening. Samples for Immunogenicity analysis (HAHA) will be taken atscreening. Pharmacodynamics Blood samples for measurement of naturalkiller (NK) cells will be collected at visits 2, 3 and 15.Pharmacokinetics Anti-EpCAM serum trough and peak levels will bemeasured at visits 2 to 6, during the treatment period every 6 to 8weeks and at the follow-up visits 16 to 18. Statistical Sample size:Methods According to sample size calculations based on Fleming'sstandard single-stage procedure but using the exact binomialdistribution (A'Hern), 24 patients evaluable for efficacy are requiredper treatment arm to provide a 85% chance (i.e. power = 80%) ofdemonstrating that the 95% one-sided confidence interval (i.e. type oneerror = 5%) for the response rate excludes 5% if the true clinicalbenefit rate is 25%. Assuming that approximately 10% of the patientswill not be evaluable with regard to efficacy (drop-outs), a total of atleast 108 patients (27 per treatment arm) should be randomized in thestudy. Statistical analysis: The primary analysis will be based on thefull analysis set. In a first step, the clinical benefit rate (patientswith stable disease ≧24 weeks + CR + PR according to RECIST (see Example1.18)) in each of the four treatment arms will be evaluated separately.A 95% one-sided confidence interval will be calculated for the clinicalbenefit rate in each treatment arm. If the lower bound of the 95%one-sided confidence interval of the response rate is larger than p₀ =5% in a treatment arm, the null hypothesis will be rejected for thistreatment arm. The cut-off point in this study is 4; meaning that assoon as 4 patients with clinical benefit have been achieved in atreatment arm the null hypothesis for the respective treatment can berejected. If all treatment arms show sufficient activity, the treatmentarms with the same dose level will be pooled and the dose levels will becompared allowing for unequal clinical benefits rates of patients withlow/moderate EpCAM expression or patients with high EpCAM expression.The comparison of the two dose levels will be conducted by means of alogistic regression model with the two factors, EpCAM expression anddose level, out of which the appropriate odds ratios for the two doselevels will be calculated. If both dose levels show sufficient activityonly in one of the patient populations (either patients withlow/moderate EpCAM expression or high EpCAM expression), these two doselevels will be compared descriptively. All secondary endpoints (efficacyand safety) will be analyzed descriptively.

Example 1.2 Summary of Non-clinical Studies with Anti-EpCAM

The efficacy of Anti-EpCAM and trastuzumab to induce ADCC was studiedusing nine breast cancer cell lines. Anti-EpCAM was capable of mediatingADCC specific lysis to the same levels as trastuzumab. There was astrong correlation between the concentration of EpCAM molecules on thecell surface and the susceptibility to ADCC using Anti-EpCAM. In allcases, maximal specific lysis was achieved at Anti-EpCAM concentrationsbelow 10 μg/mL, which is the minimal trough concentration targeted inpatients.

Anti-EpCAM showed an excellent local tolerance in rabbits at theintended intravenous route of administration, no macroscopic and onlyminor microscopic changes were observed.

Example 1.3 Rationale for Dose Selection

Based on preclinical experiments, serum trough levels of 10 μg/mL areexpected to be effective for anti-tumor activity of Anti-EpCAM. However,it cannot be ruled out that higher doses might be more effective.Therefore, a second dose, calculated to achieve serum trough level of 30μg/mL, will be evaluated in this study.

The doses intended in this study do not exceed the highest dosesadministered to patients in the phase I study. Loading phases andmaintenance phases have been calculated using pharmacokinetic modelingto achieve targeted trough serum concentrations within a short period oftime and to avoid maximum plasma concentrations that would exceed theones assessed in the phase I study.

Example 1.4 Known Benefits

Preclinical data suggest that patients with metastatic breast cancer andother tumors may benefit from a delay of progression (stable disease) byelimination of tumor cells, which express EpCAM, via Anti-EpCAM.

Example 1.5 Clinical Study Objectives

Primary Objective

-   -   To evaluate the clinical benefit of two different doses of        Anti-EpCAM in patients with EpCAM positive, metastatic breast        cancer        Secondary Objectives    -   To evaluate other response parameters for two different doses        for Anti-EpCAM    -   To evaluate the safety and tolerability of two different doses        of Anti-EpCAM    -   To determine the pharmacokinetics of two different doses of        Anti-EpCAM    -   To evaluate pharmacodynamics of two different doses of        Anti-EpCAM    -   To determine the pharmacokinetics of Anti-EpCAM in patients with        metastatic breast cancer    -   To evaluate the pharmacodynamics of Anti-EpCAM (NK cells)

Example 1.6 Investigational Plan

Study Endpoints; Primary EndpointClinical benefit rate (SD+PR+CR) at 24weeks

The clinical benefit rate is defined as the proportion of patients withstable disease (SD)+partial response (PR)+complete response (CR),according to RECIST (see Example 1.18).

Study Endpoints; Secondary Endpoints

-   -   Clinical benefit rate (SD+PR+CR) at 12 weeks    -   Best overall tumor response rate (OTR)    -   Duration of response/time to progression    -   Incidence of adverse events and laboratory abnormalities    -   Serum concentrations of Anti-EpCAM    -   Number of peripheral natural killer (NK) cells        Overall Study Design

This is an open-label, multicenter, randomized, parallel group, phase IIstudy investigating the efficacy and safety of two treatment doses ofAnti-EpCAM in patients with low/moderate or high EpCAM expression over24 weeks of therapy.

A total of 112 patients have been enrolled in the study. After thescreening period, the patients meeting all illegibility criteria will berandomized within each EpCAM stratum to one of two treatment groups: alow dose group and a high dose group. Anti-EpCAM will be administered asa 60 minutes intravenous (i.v.) infusion weekly during the loading phase(Day 1, Day 8 and Day 15) and every second week thereafter, for a totalof 24 weeks or until disease progression. Patients were stratifiedaccording to the level of their EpCAM expression. Here, there were twogroups: those with low/moderate EpCAM expression, and those with highEpCAM expression. Table 2 shows an overview of this additionalstratification.

TABLE 2 Stratified treatment groups and Anti-EpCAM doses TreatmentGroups EpCAM Expression Anti-EpCAM Dosing Group I Moderate EpCAM 2 mg/kgAnti-EpCAM i.v., expression on primary tumor every two weeks Group IIModerate EpCAM 6 mg/kg Anti-EpCAM i.v., expression on primary tumorevery two weeks Group III High EpCAM expression on 2 mg/kg Anti-EpCAMi.v., primary tumor every two weeks Group IV High EpCAM expression on 6mg/kg Anti-EpCAM i.v., primary tumor every two weeks

Patients will be monitored every 6 weeks until week 24 and every 8 weeksthereafter (during follow-up study), by a clinical assessment andlaboratory tests. Further evaluations will be performed for theassessment of tumor response including thoracic CT scan or chest X-ray,abdominal CT scan or MRI, and bone scintigraphy if bone lesions weredetected at screening.

The efficacy will be evaluated for each group, and the duration ofresponse/time to progression will be compared to the expected values inthis patient population. Tumor response as measured by RECIST (seeExample 1.18) criteria will be assessed and used for statisticalanalysis.

The primary endpoint of the study is the clinical benefit rate(CR+PR+SD) at week 24 in each of the four groups. The clinical benefitrate (SD+PR+CR) at week 12, best overall tumor response rate (OTR),duration of response/time to progression will be evaluated as secondaryendpoints. The safety and tolerability results will be compared betweentreatment groups at week 24.

If it becomes evident at any time during the study that one of the dosesis preferable for efficacy or tolerability reasons, the study protocolwill be amended accordingly.

Patients for whom SD, PR or CR is documented after 24 weeks of therapyand in whom no unacceptable toxicity (and no treatment interruptiongreater than 4 weeks) was reported, will be offered to participate in afollow-on study with continued Anti-EpCAM therapy. Parameters such aslong-term tolerability, clinical progression and overall survival willbe evaluated.

Example 1.7 Clinical Study Population

Inclusion Criteria

-   -   1. A patient will be eligible for study participation only if        all of the following criteria apply:    -   2. Histologically confirmed metastatic breast cancer with        positive EpCAM expression in the archived tissue samples        determined by immunohistochemistry at screening    -   3. Presence of at least one lesion (i.e. metastasis) measurable        in at least one dimension (according to RECIST (see Example        1.18))    -   4. Life expectancy≧12 months    -   5. ECOG performance status 0-1    -   6. Age≧18 years    -   7. Ability to understand and willingness to sign a written        informed consent        Exclusion Criteria

A patient will not be eligible to participate in this study if any ofthe following criteria apply:

-   -   1. Any other treatment to be recommended/preferred at time of        inclusion as per Investigator's assessment    -   2. History of CNS metastases    -   3. Indication for trastuzumab (Herceptin®) treatment as per        Investigator's assessment    -   4 Immunotherapy, radiation, chemotherapy or any other anticancer        therapy within 4 weeks prior to start of therapy except:        -   Localized radiotherapy that started prior to visit 1 (target            lesion cannot be within area of irradiation, and            radiotherapy should not be expected to result in marrow            suppression as defined in exclusion criterion 6)        -   Hormonal treatment under which patient was progressive and            which is terminated prior start of study treatment    -   5. Any investigational product within 4 weeks prior to the start        of therapy    -   6. Abnormal organ or bone marrow function defined as follows:        -   Hemoglobin concentration≦90 g/L of 9.0 g/dL        -   Leukocytes<3×10⁹/L (3000/mm³)        -   Platelet count<100×10⁹/L (100,000/mm³)        -   AST(SGOT) or ALT(SGPT)>2×upper limit of normal (ULN) (>5×ULN            if liver metastases present)        -   Serum creatinine>1.5×ULN        -   Serum lipase>1.5×ULN        -   Serum amylase>1.5×ULN    -   7. History of malignancy other than breast cancer within 5 years        prior to start of therapy, with the exception of basal cell        carcinoma of the skin or carcinoma in situ of the cervix    -   8. Any other concurrent disease or medical condition that is        deemed to interfere with the conduct of the study as judged by        the investigator    -   9. Anticipated need, or regular use within 4 weeks prior to        start of therapy, of immunosuppressive agents such as systemic        corticosteroids    -   10. Known infection with human immunodeficiency virus (HIV)        and/or infection with hepatitis B virus (HbsAg positive) or        hepatitis C virus (anti-HCV positive)    -   11. Pregnant or nursing women, or women of childbearing        potential not willing to use an effective form of contraception        during participation in the study and at least 3 months        thereafter    -   12. Known hypersensitivity to immunoglobulins or to any other        component of the study drug formulation

Example 1.8 Clinical Trial Material Preparation

The investigational product Anti-EpCAM for clinical trial use issupplied as a solution in GMP quality, for example comprising 10 mg/mLAnti-EpCAM in isotonic phosphate buffer and is stored between +2 and +8°C.

The amount of Anti-EpCAM for the preparation of the final solution forinfusion to a patient is calculated based on the patient's body weightand treatment group (see above Table 2).

Anti-EpCAM will be diluted in 500 mL 0.9% sodium chloride solution in aclean, sterile environment (laminar flow hood). To mix the finalAnti-EpCAM solution for infusion, the bag should be inverted gently toavoid foaming Both the concentrate and final solution for infusion arefor single use only.

Treatment Assignment Procedure

Randomization will take place as close as possible to the start ofAnti-EpCAM therapy. All eligibility criteria must be met at the time ofrandomization. A centralized randomization procedure will be provided bymeans of an ICRS (Interactive Computer Response System). Theinvestigator has to login with an individual identification number andpassword onto a secured website where she/he has to provide basicpatient data (patient number, data of screening, date of birth, EpCAMtest results) in order to receive immediate response regarding treatmentassignment. The randomization procedure does not stratify the treatmentassignment by center but by EpCAM expression to ensure balanceddistribution of high and low dose treatments in both EpCAM expressionstrata. As soon as the requested number of patients has been achieved ina treatment arm no further patients can be randomized to this respectivearm.

The solution should be administered to the patient intravenously over 60minutes, at a flow rate of 500 mL/h

Example 1.9 Treatment

Treatment Schedule

Each patient will receive a total of 14 infusions of Anti-EpCAM over 24weeks of therapy unless disease progression or treatment-limitingtoxicity occurs. The Anti-EpCAM solution for infusion will beadministered to the patient intravenously over 60 minutes, weekly duringthe loading phase and every second week during the maintenance phase.Prior to Visit 2 (Day 1), the patients will be randomized within eachEpCAM expression stratum to one of the following treatment groups:

-   -   Groups I and III (low dose): loading phase of 2 mg Anti-EpCAM/kg        body weight weekly (week 1, 2 and 3), followed by 11 maintenance        doses of 2 mg Anti-EpCAM/kg body weight every second week    -   Groups II and IV (high dose): loading phase of 6 mg        Anti-EpCAM/kg body weight at weekly (week 1, 2 and 3), followed        by 11 maintenance doses of 6 mg Anti-EpCAM/kg body weight every        second week

Randomization procedure will be centralized and stratified according tothe EpCAM test result performed at screening. An interactive computerresponse system will be used to register EpCAM results and patient data.Upon registration of the data treatment assignment will be given for thepatient. Calculation of the individual doses for a patient andpreparation of the final solution for infusion will be performed at eachcenter.

Patients for whom SD, PR or CR is documented after 24 weeks of therapyand in whom no unacceptable toxicity (and no treatment interruptiongreater than 4 weeks) was reported will be offered to participate in afollow-up study with continued Anti-EpCAM therapy.

Treatment Discontinuation for Adverse Events

The study medication may be interrupted or discontinued, or the dose maybe reduced for patients in the high dose group, according to theseverity and causality of the adverse event.

Should the investigator have compelling evidence that the adverse eventis not caused by the study medication, the treatment should becontinued. However, if the relationship of the adverse event to thestudy medication cannot be excluded, doses should be modified.

Patient Discontinuation Criteria

Treatment with the investigational product should be discontinued i.a.in the event of any of the following:

-   -   Disease progression, as defined by RECIST (see Example 1.18)    -   Withdrawal of patient's consent    -   Patient or investigator not compliant with the study protocol    -   Progression of a medical condition which in the opinion of the        investigator should preclude further participation of the        patient in the study    -   Administration of non-permitted concomitant medication(s)    -   Investigator's decision that a change of therapy is in the        patient's best interest    -   Occurrence of a Grade 3 adverse event if        -   the adverse event is seen as clinically significant by the            investigator and        -   does not resolve to ≦grade 2 prior to the next            administration and        -   is at least possibly related to the study medication.    -   Occurrence of a Grade 4 adverse event Dose interruption for more        than 4 weeks    -   Occurrence of any adverse event which makes discontinuation        desirable or necessary in the investigator's and/or the        patient's opinion.

Prior to discontinuation of a patient, all examinations scheduled forthe safety follow-up should also be performed to allow for theevaluation of the study endpoints (see Example 1.11).

Concomitant Medication

All concomitant medication should be recorded in the Case Report Form(CRF). The following medication and therapies are not allowed during thewhole study period:

-   -   Any anti-tumor therapy other than the investigational product        such as:        -   Hormonal therapy        -   Biological therapy        -   Chemotherapy        -   Radiation therapy (Exception: Localized radiotherapy that            started prior to visit 1 (target lesion cannot be within            area of irradiation, and radiotherapy should not be expected            to result in marrow suppression as defined in exclusion            criterion 6) Pre-menopausal patients treated with LHRH            analogs (Hormone receptor+) could continue this treatment            during the study provided that they progressed on LHRH            analogs before study entry.    -   Chronic systemic high-dose corticosteroid therapy and other        immunosuppressive therapies    -   Any other investigational agent

If required, supportive therapy should be administered as medicallyneeded in accordance with standard practice and should be recorded inthe CRF. Therapy with bisphosphonates is allowed for patients with bonemetastasis at study entry.

Example 1.10 Assessments

Assessment of Eligibility/Efficacy

Informed Consent: Written informed consent must be obtained from eachpatient before any study specific procedure.

Inclusion/Exclusion: Assessment of the patient's eligibility should beperformed as outlined above in Example 1.7—Clinical Study Population,including review of all laboratory measurements performed for screening.

Medical History/Current Medical Conditions: General and disease specificmedical history including a history of past and current medicalconditions; full history of the course of the patient's cancer,including tumor stage, other prognostic markers, information on prioranti-tumor therapies will be recorded at screening.

Concomitant Medication: All concomitant medications will be recordedthroughout the study.

Hepatitis B and C Testing: Hepatitis B surface antigen (HBsAg) andHepatitis C virus (HCV) antibody testing will be performed during thescreening period to exclude an active infection with hepatitis B or Cviruses.

Tumor Assessment: Tumor response will be defined according to the RECIST(see Example 1.18) criteria

-   -   Bone Scintigraphy: Bone scintigraphy will be performed to assess        the presence of bone metastasis at screening and at the final        visit. Further scans to be performed if bone metastases are        present at screening or if clinically indicated (e.g. occurrence        of pain and elevation of alkaline phosphatase).    -   Thoracic Computerized-Tomography (CT) Scan or Chest X-ray:        Thoracic CT or chest X-ray will be performed to document the        presence of distant metastasis at screening and to evaluate        response regularly during the treatment phase and at the final        visit.    -   Abdominal CT Scan or MRI: Abdominal CT scan or MRI will be        performed to assess the presence of distant metastasis at        screening and to evaluate response regularly during treatment        and at the final visit.        Assessment of Safety

Adverse Events: Adverse events (AEs) occurring during the treatmentperiod and until 4 weeks after the last Anti-EpCAM infusion will berecorded. During the screening period, AEs related to study proceduresshould also be reported.

Serious Adverse Events: All serious adverse events (SAE) will berecorded during the entire study period, including the screening and thefollow-up periods.

Physical Examination: A complete physical examination of all bodysystems including vitals signs will be performed at screening, atregular intervals during the treatment phase and at the final visit. Asymptom directed physical examination will be performed throughout thestudy as appropriate, and all clinically relevant findings will bedocumented. On infusion days, physical examination should be performedbefore Anti-EpCAM infusion.

Vital Signs Monitoring: Body temperature (oral or tympanic), heart rateand blood pressure (systolic/diastolic) will be measured throughout thestudy as follows:

-   -   prior to infusion    -   every 15 minutes during infusion at visits 2 to 6    -   every hour for up to 4 hours after end of the infusion at visits        2 to 6

ECOG Score: Assessment of the patient's performance status will beperformed using the Eastern Cooperative Oncology Group (ECOG) score (seeTable 6 in Example 1.19).

Electrocardiogram (ECG): Standard 12-lead ECG will be performed atscreening, during the treatment phase and at the final visit. For eachECG, two printouts should be obtained, one for documentation at the siteand one for evaluation by a central cardiologist.

Safety Laboratory Evaluations: Blood samples for safety laboratoryevaluations will be taken at each study visit in the morning (before theinfusion on infusion days) and the following analyses will be performed:

-   -   Clinical chemistry: Aspartate transaminase (AST), alanine        transaminase (ALT), γ-glutamyl transferase (GGT), alkaline        phosphatase (AP), lactate dehydrogenase (LDH), total bilirubin,        total protein, creatinine, urea, uric acid, glucose, calcium,        sodium, potassium, chloride, phosphate, amylase, lipase,        albumin, C-reactive protein (CRP).    -   Hematology: Red blood cell count (RBC), hemoglobin, hematocrit,        white blood cell count (WBC), differential blood count, and        platelet count.    -   Coagulation: Prothrombin time (PT, international normalized        ratio [INR]), partial thromboplastin time (PTT) and fibrinogen.

Urinalysis: Presence of glucose, protein and blood in urine will beassessed by dipstick at each visit. On the days of Anti-EpCAM infusion,urinalysis should be performed before Anti-EpCAM infusion.

Pregnancy Test: A pregnancy test (β-human chorionic gonadotropine[(β-HCG]) will be performed at screening and at the final visit in allwomen of childbearing potential.

Immunogenicity: Blood samples for the assessment of Anti-EpCAMimmunogenicity will be taken at screening, at weeks 6 and 24 and thelast two follow-up visits 17 and 18.

Pharmacodynamic Assessments

Natural Killer Cells: The number of natural killer (NK) cells will bemeasured by Fluorescence-Activated Cell Sorter (FACS).

Pharmacokinetic Assessments

Anti-EpCAM serum trough and peak levels will be measured at visits 2 to6, during the treatment period every 6 to 8 weeks and the last threefollow-up visits 16 to 18.

Analysis of EpCAM expression

EpCAM expression will be assessed at screening, using the patientsarchived tumor material. EpCAM expression will be determined byimmunohistochemistry in a central laboratory.

Only patient with EpCAM results, which are low/moderate or high canproceed with all screening procedures. Patients with negative EpCAM testresult do not meet eligibility criteria and will be considered asscreening failures.

If during the study a biopsy is performed (e.g. in case of new detectedmetastasis), tumor tissue should also be collected for analysis of EpCAMexpression.

Example 1.11 Visit Schedule

The calculation of all study visits is based on baseline (Day 1),defined as the day of the first dose of Anti-EpCAM infusion.

Screening Period (Day-28 to Day-1)

Only those patients who have met all eligibility criteria will beassigned a patient number (see Example 1.8—Treatment AssignmentProcedure).

All screening evaluations must be performed within 28 days of the firstadministration of the investigational product (Day 1). All results,including the EpCAM expression test should be available before a patientis declared eligible for study participation. Once the results from thelaboratory evaluations are available and the patient meets alleligibility criteria, the investigator should proceed with thecentralized randomization procedure (ICRS).

Treatment Period (Visits 2 to 15)

During the treatment period, the following procedures and assessmentsshould be performed:

-   -   Vital Signs    -   Physical Examination    -   Safety Laboratory Evaluation    -   Urinalysis    -   Pharmacokinetics    -   NK cells    -   Immunogenicity    -   Concomitant Medication    -   Adverse Events and Serious Adverse Events    -   Chest X-ray/CT scan    -   Abdominal CT scan or MRI    -   Bone scintigraphy (if bone lesions detected at screening)    -   ECG (only at visit 6 and 15)        Safety Follow-up Period and Final PK/PD Assessment (Visits        16-18)

Safety follow-up visits will be performed two and four weeks after theend of therapy. Visit 17 is the final safety follow-up visit.

The last study visit (Visit 18) is the Final PK/PD Assessment 12 weeksafter the end of therapy.

End of Study Visit (Visit 17/Final)

The end of study visit should be performed at the last follow-up visitor at any time if the patient is prematurely discontinued from thestudy.

-   -   Complete Physical Examination    -   Vital Signs    -   12-lead ECG    -   ECOG    -   Safety laboratory evaluation    -   Urinalysis    -   Pregnancy test    -   Chest X-ray/CT scan    -   Bone scintigraphy    -   Abdominal CT scan or MRI    -   Pharmacokinetics    -   Immunogenicity    -   Concomitant Medication    -   Adverse Events and Serious Adverse Events        Efficacy Follow-up Period

Patients will be followed for disease progression, other cancertreatment and survival every 3 months until 1 year after the end ofparticipation in the study. Patients with stable disease, partial orcomplete response at week 24 will have the opportunity to enter into anopen-label study, in which evaluation of disease progression and overallsurvival will be performed.

Example 1.12 Sample Storage

Serum samples for immunogenicity, NK cell count and PK will be stored atthe study site frozen at −20° C.

Example 1.13 Safety Considerations

The investigator is responsible for the detection and documentation ofevents meeting the definition of an adverse event (AE) or a seriousadverse event (SAE). This includes the evaluation of its seriousness,its severity, and the causal relationship to the investigational productand/or concomitant therapy.

Example 1.14 Adverse Events, Serious Adverse Events

Adverse Events

The following events, detected or diagnosed during or afteradministration of the investigational medicinal product, are adverseevents:

-   -   Aggravation of a pre-existing illness or permanent disorder,    -   Increase in frequency or intensity of a pre-existing episodic        event or condition,    -   Signs or symptoms detected or diagnosed after administration of        the investigational medicinal product, even if they may have        occurred or existed prior to study participation,    -   Marked hematological and other laboratory abnormalities and any        events that led to an intervention, including withdrawal of test        drug/investigational product treatment, dose reduction, or        significant additional concomitant therapy.        Serious Adverse Event

A serious adverse event (experience) or reaction is any untoward medicaloccurrence or effect that at any dose:

-   -   Results in death, (1)    -   Is life-threatening, (2)    -   Requires hospitalization or prolongation of existing inpatient's        hospitalization,    -   Results in persistent or significant disability or incapacity,    -   Is a congenital anomaly or birth defect.

Assessment of Severity: The severity (or intensity) of AEs is evaluatedaccording to the grading scale provided in the Cancer Therapy EvaluationProgram, Common Terminology Criteria for Adverse Events (CTCAE), version3.0.

Example 1.15 Statistical Analysis

Statistical Methods and Determination of Study Variables

The aim of this study is to determine efficacy and safety of Anti-EpCAM.Therefore, a randomized phase II trial with two arms (low and high doseof Anti-EpCAM) is performed, in which each treatment arm shall be deemedas a standard single-stage phase II study. The decision whether atreatment proofs to show sufficient activity will be made for eachtreatment arm separately. Sample size estimation and primary statisticalanalysis are based on standard single-stage single-arm phase II designs.

Table 3 lists the measurements and/or data management processes thatwill be carried out to determine the primary and secondary studyendpoints.

TABLE 3 Primary and secondary study objectives, derived variables andmethod of measurement. Objective Endpoint Measurement Primary Toevaluate the Clinical benefit Proportion of clinical benefit of rate atweek 24 patients with SD, two different doses PR and CR at of Anti-EpCAMin week 24 patients with EpCAM positive, metastatic breast cancerSecondary To evaluate Clinical benefit Proportion of other response rateat week 12 patients with SD, parameters for PR and CR at two differentdoses week 12 and 48 for Anti-EpCAM Best overall Proportion of tumorresponse patients achieving rate (OTR) a PR/CR throughout study periodDuration of Median time to response/time to disease progressionprogression calculated using Kaplan-Meier- methodology To evaluate theIncidence of Proportion of safety and tolerability adverse eventspatients with events: of two different doses and laboratory number,intensity and of Anti-EpCAM abnormalities relationship to studytreatment (according to investigator) of laboratory abnormalities,clinical adverse events and serious adverse events To determine theSerum Serum pharmacokinetics concentrations concentrations of Anti- oftwo different of Anti-EpCAM EpCAM at defined doses of Anti- time-pointsEpCAM To evaluate Number of % Increase/decrease pharmacodynamicsperipheral in NK-cell of two different natural killer counts over timedoses of Anti-EpCAM (NK) cells

Statistical Hypothesis: The following assumptions are made for thestatistical hypothesis of the study:

The background clinical benefit rate for best supportive care isestimated to be ≦5% (p₀)). The future use of Anti-EpCAM in the describedpatient population (positive EpCAM expression) would be of considerableinterest if the true clinical benefit rate (π) is ≧25% (p₁).

For each treatment arm (low or high dose group within the low/moderateand the high EpCAM strata), the following hypotheses apply:

-   -   H_(0ij)(π≦p_(0ij)): p_(ij)≦5% (background clinical benefit        probability)    -   H_(1ij)(π≧p_(1ij)): p_(ij)≧25% (clinical benefit probability of        interest for Anti-EpCAM) with p_(i) being the clinical benefit        rate observed among the low/moderate EpCAM expression strata        (j=1) and high EpCAM expression strata patients (j=2) treated        with dose level i, with i=1 denoting the low dose level and i=2        indicating the high dose level.

Level of Significance, Multiple Comparisons and Multiplicity: A type Ierror of 5% and a type II error of 15% (power of 85%) are considered asadequate to determine the clinical benefit rates. No significance levelshave to be adjusted neither due to multiple comparisons nor due tomultiplicity (no confirmatory comparison between treatment arms will beperformed).

Determination of Sample Size: The sample size estimation is based onFleming's standard single-stage procedure but using the exact binomialdistribution (A′Hern (2001). Statistics in Medicine 20: 859-66), and notthe normal approximation to the binomial distribution (Fleming (1982).Biometrics 38: 143-51). This approach is preferred because normalapproximations are incorrect for small sample sizes, and sample sizesand cut-off points based on exact distributions have the advantage thatthe calculated confidence intervals do not include p₀ if the cut-offpoint has been achieved. Therefore, sample size tables provided byA'Hern (A'Hern (2001). Statistics in Medicine 20: 859-66) are employedfor this study.

According to these calculations, 24 patients evaluable for efficacy arerequired per arm for the study to have a 85% chance (i.e. power=85%) ofdemonstrating that the 95% one-sided confidence interval (i.e. type oneerror=5%) for the response rate excludes 5% (p₀) if the true overallresponse rate over 24 weeks is 25% (p₁).

Assuming that approximately 10% of the patients will not be evaluablewith regard to efficacy (drop-outs), a total of at least 108 patients(54 in the low/moderate EpCAM strata with 27 of those treated with thelow dose and 27 with the high dose and 54 in the high EpCAM expressionstrata with 27 of those treated with low dose and 27 with the high dose)should be randomized into the study.

Planned Analyses

Primary and secondary variables will be evaluated exploratively. Allrelevant data on patients (data from CRF, laboratory data) will beanalyzed descriptively grouped by treatment arm and visits.

Individual patient data will be presented in listings (sorted bytreatment arm and patient number). All data collected in the CRF andincluded in the database will be listed.

Demographic and Other Baseline Characteristics: Demographics and otherbaseline characteristics will be summarized in total and by treatmentarm by means of summary statistics (number of patients, mean, standarddeviation, minimum, median, maximum) for continuous variables and byabsolute and relative frequencies for categorical variables. Baselinecharacteristics are defined as all results of the examinations performedprior to the first Anti-EpCAM administration.

Planned Analyses for Primary Endpoint: The primary analysis will bebased on the full analysis set, an analysis based on the per-protocolset will be performed as a sensitivity analysis. Primary endpoint of thestudy is the clinical benefit rate (patients with stable disease+CR+PRaccording to RECIST (see Example 1.18))

In a first step, the clinical benefit rate in each treatment arm will beevaluated separately. A 95% one-sided confidence interval will becalculated for the clinical benefit rate in each treatment arm. If thelower bound of the 95% one-sided confidence interval of the responserate is larger than p₀=5% in a treatment arm, the null hypothesis willbe rejected for this treatment arm. The cut-off point in this study is4, meaning that as soon as 4 patients with clinical benefit have beenachieved in a treatment arm the null hypothesis for the respectivetreatment arm can be rejected Further analysis of the primary endpointwill include:

-   -   If all treatment arms show sufficient activity, the treatment        arms with the same dose level will be pooled and the dose levels        will be compared allowing for unequal clinical benefit rates for        patients with low/moderate and high EpCAM expression. The        comparison of the two dose levels will be conducted by means of        a logistic regression model with the two factors EpCAM        expression and dose level, out of which the appropriate odds        ratios for the two levels will be calculated    -   If both dose levels show sufficient activity only in one of the        patient populations (either with low/moderate EpCAM expression        or with high EpCAM expression) these two dose levels will be        compared descriptively.

Planned Analyses for Secondary Endpoints: Best overall tumor responserate at week 24 will be evaluated for each treatment arm and in the sameway as described for the primary endpoint.

Adverse events will be summarized overall by treatment arm and by doselevel, grouped by their primary system organ class, high level term,preferred term and severity.

No statistical comparison of the overall incidence of adverse eventswill be done between the treatment arms and dose levels.

Data will be summarized for all sample days overall, by treatment armand by dose level. Absolute changes from baseline values for allpost-baseline sample days will be summarized overall, by treatment armand by dose level. Shift tables from baseline will be produced forurinalysis data overall, by treatment arm and by dose level.

Pharmacodynamic parameters will be analyzed descriptively by presentingsummary statistics (mean, standard deviation, minimum, median andmaximum) of raw data and changes from baseline for all study daysoverall, by treatment arm and by dose level.

Administrative Data Review

For the purpose of deciding on further strategies for subsequent studieswith Anti-EpCAM, an administrative analysis of the best overall responserate will be performed,

-   -   after 70 patients received at least one infusion and have passed        Visit 9/week 12 or prematurely discontinued the study and    -   data regarding the best overall response rate is considered to        be reasonably clean

Recruitment will not be stopped for this administrative analysis.

Example 1.16 Quality Control and Quality Assurance

Prior to enrolment of patients at a study site, specific regulatorydocuments must be available, such as independent ethics committee (IEC)approval and curricula vitae for investigator and study staff.

The study will be monitored by a qualified and appropriately trainedperson appointed by Study Sponsor.

Example 1.17 Legal and Ethical Requirements

This study will be conducted in accordance with the ICH HarmonizedTripartite Guideline Guidelines for Good Clinical Practice and allapplicable laws and regulations, including the Declaration of Helsinki,June 1964, as modified by the 48^(th) World Medical Association,Somerset West, Republic of South Africa, October 1996.

Patient Information and Informed Consent

The process for obtaining patient informed consent will be in accordancewith all applicable regulatory requirements. Prior to including anypatient in the clinical study, his/her free and expressed informedconsent must be obtained in writing.

Example 1.18 RECIST Criteria

The following is a summary of the RECIST criteria used throughout theforegoing study.

Eligibility

Only patients with measurable disease at baseline should be included inprotocols where objective tumor response is the primary endpoint.

Measurable disease—the presence of at least one measurable lesion. Ifthe measurable disease is restricted to a solitary lesion, itsneoplastic nature should be confirmed by cytology/histology.

Measurable lesions—lesions that can be accurately measured in at leastone dimension with longest diameter ≧20 mm using conventional techniquesor ≧10 mm with spiral CT scan.

Non-measurable lesions—all other lesions, including small lesions(longest diameter <20 mm with conventional techniques or <10 mm withspiral CT scan), i.e., bone lesions, leptomeningeal disease, ascites,pleural/pericardial effusion, inflammatory breast disease, lymphangitiscutis/pulmonis, cystic lesions, and also abdominal masses that are notconfirmed and followed by imaging techniques.

All measurements should be taken and recorded in metric notation, usinga ruler or calipers. All baseline evaluations should be performed asclosely as possible to the beginning of treatment and never more than 4weeks before the beginning of the treatment.

The same method of assessment and the same technique should be used tocharacterize each identified and reported lesion at baseline and duringfollow-up.

Clinical lesions will only be considered measurable when they aresuperficial (e.g., skin nodules and palpable lymph nodes). For the caseof skin lesions, documentation by color photography, including a rulerto estimate the size of the lesion, is recommended.

Methods of Measurement

CT and MRI are the best currently available and reproducible methods tomeasure target lesions selected for response assessment. Conventional CTand MRI should be performed with cuts of 10 mm or less in slicethickness contiguously. Spiral CT should be performed using a 5 mmcontiguous reconstruction algorithm. This applies to tumors of thechest, abdomen and pelvis. Head and neck tumors and those of extremitiesusually require specific protocols.

Lesions on chest X-ray are acceptable as measurable lesions when theyare clearly defined and surrounded by aerated lung. However, CT ispreferable.

When the primary endpoint of the study is objective response evaluation,ultrasound (US) should not be used to measure tumor lesions. It is,however, a possible alternative to clinical measurements of superficialpalpable lymph nodes, subcutaneous lesions and thyroid nodules. US mightalso be useful to confirm the complete disappearance of superficiallesions usually assessed by clinical examination.

The utilization of endoscopy and laparoscopy for objective tumorevaluation has not yet been fully and widely validated. Their uses inthis specific context require sophisticated equipment and a high levelof expertise that may only be available in some centers. Therefore, theutilization of such techniques for objective tumor response should berestricted to validation purposes in specialized centers. However, suchtechniques can be useful in confirming complete pathological responsewhen biopsies are obtained.

Tumor markers alone cannot be used to assess response. If markers areinitially above the upper normal limit, they must normalize for apatient to be considered in complete clinical response when all lesionshave disappeared.

Cytology and histology can be used to differentiate between PR and CR inrare cases (e.g., after treatment to differentiate between residualbenign lesions and residual malignant lesions in tumor types such asgerm cell tumors).

Baseline documentation of “Target” and “Non-Target” lesions

All measurable lesions up to a maximum of five lesions per organ and 10lesions in total, representative of all involved organs should beidentified as target lesions and recorded and measured at baseline.

Target lesions should be selected on the basis of their size (lesionswith the longest diameter) and their suitability for accurate repeatedmeasurements (either by imaging techniques or clinically).

A sum of the longest diameter (LD) for all target lesions will becalculated and reported as the baseline sum LD. The baseline sum LD willbe used as reference by which to characterize the objective tumor.

All other lesions (or sites of disease) should be identified asnon-target lesions and should also be recorded at baseline. Measurementsof these lesions are not required, but the presence or absence of eachshould be noted throughout follow-up.

Response Criteria

Evaluation of Target Lesions

-   -   Complete Response (CR): Disappearance of all target lesions    -   Partial Response (PR): At least a 30% decrease in the sum of the        LD of target lesions, taking as reference the baseline sum LD    -   Progressive Disease (PD): At least a 20% increase in the sum of        the LD of target lesions, taking as reference the smallest sum        LD recorded since the treatment started or the appearance of one        or more new lesions    -   Stable Disease (SD): Neither sufficient shrinkage to qualify for        PR nor sufficient increase to qualify for PD, taking as        reference the smallest sum LD since the treatment started

Evaluation of Non-target Lesions

-   -   Complete Response (CR): Disappearance of all non-target lesions        and normalization of tumor marker level    -   Incomplete Response/Persistence of one or more non-target        lesion(s) or/and    -   Stable Disease (SD): maintenance of tumor marker level above the        normal limits    -   Progressive Disease (PD): Appearance of one or more new lesions        and/or unequivocal progression of existing non-target lesions        (1)

-   (1) Although a clear progression of “non target” lesions only is    exceptional, in such circumstances, the opinion of the treating    physician should prevail and the progression status should be    confirmed later on by the review panel (or study chair).    Evaluation of Best Overall Response

The best overall response is the best response recorded from the startof the treatment until disease progression/recurrence (taking asreference for PD the smallest measurements recorded since the treatmentstarted). In general, the patient's best response assignment will dependon the achievement of both measurement and confirmation criteria

Target lesions New lesions Non-Target lesions Overall response CR CR NoCR CR Incomplete No PR response/SD PR Non-PD No PR SD Non-PD No SD PDAny Yes or No PD Any PD Yes or No PD Any Any Yes PD

Patients with a global deterioration of health status requiringdiscontinuation of treatment without objective evidence of diseaseprogression at that time should be classified as having “symptomaticdeterioration”. Every effort should be made to document the objectiveprogression even after discontinuation of treatment.

In some circumstances it may be difficult to distinguish residualdisease from normal tissue. When the evaluation of complete responsedepends on this determination, it is recommended that the residuallesion be investigated (fine needle aspirate/biopsy) to confirm thecomplete response status.

Confirmation

The main goal of confirmation of objective response is to avoidoverestimating the response rate observed. In cases where confirmationof response is not feasible, it should be made clear when reporting theoutcome of such studies that the responses are not confirmed.

To be assigned a status of PR or CR, changes in tumor measurements mustbe confirmed by repeat assessments that should be performed no less than4 weeks after the criteria for response are first met. Longer intervalsas determined by the study protocol may also be appropriate.

In the case of SD, follow-up measurements must have met the SD criteriaat least once after study entry at a minimum interval (in general, notless than 6-8 weeks) that is defined in the study protocol.

Duration of Overall Response

The duration of overall response is measured from the time measurementcriteria are met for CR or PR (whichever status is recorded first) untilthe first date that recurrence or PD is objectively documented, takingas reference for PD the smallest measurements recorded since thetreatment started.

Duration of Stable Disease

SD is measured from the start of the treatment until the criteria fordisease progression are met, taking as reference the smallestmeasurements recorded since the treatment started.

The clinical relevance of the duration of SD varies for different tumortypes and grades. Therefore, it is highly recommended that the protocolspecify the minimal time interval required between two measurements fordetermination of SD. This time interval should take into account theexpected clinical benefit that such a status may bring to the populationunder study.

Response Review

For trials where the response rate is the primary endpoint it isstrongly recommended that all responses be reviewed by an expert(s)independent of the study at the study's completion. Simultaneous reviewof the patients' files and radiological images is the best approach.

Reporting of Results

All patients included in the study must be assessed for response totreatment, even if there are major protocol treatment deviations or ifthey are ineligible. Each patient will be assigned one of the followingcategories: 1) complete response, 2) partial response, 3) stabledisease, 4) progressive disease, 5) early death from malignant disease,6) early death from toxicity, 7) early death because of other cause, or9) unknown (not assessable, insufficient data). All of the patients whomet the eligibility criteria should be included in the main analysis ofthe response rate. Patients in response categories 4-9 should beconsidered as failing to respond to treatment (disease progression).Thus, an incorrect treatment schedule or drug administration does notresult in exclusion from the analysis of the response rate. Precisedefinitions for categories 4-9 will be protocol specific.

All conclusions should be based on all eligible patients.

Subanalyses may then be performed on the basis of a subset of patients,excluding those for whom major protocol deviations have been identified(e.g., early death due to other reasons, early discontinuation oftreatment, major protocol violations, etc.). However, these subanalysesmay not serve as the basis for drawing conclusions concerning treatmentefficacy, and the reasons for excluding patients from the analysisshould be clearly reported. The 95% confidence intervals should beprovided.

Example 1.19 Overview of ECOG Performance Status

The ECOG performance status scale is disclosed in Oken, M. M. et al.(1982) Am J Clin Oncol 5:649-655.

Example 2 Administrative Data Review for Phase II Clinical Study ofAnti-EpCAM Example 2.1 Summary and Introduction to Study

The clinical study was performed as described above in Example 1. Theresults of this study follow now in Example 2.

Methodology:

A randomized, open-label, multicenter, parallel group, phase II study.The study was designed to evaluate the efficacy and safety of Anti-EpCAMover 24 weeks of therapy at two different doses with positive EpCAMtesting. The central randomization process was stratified according tothe EpCAM test results performed at screening. Upon registration in oneof the EpCAM strata, patients were randomly assigned to either the lowdose treatment group or the high dose treatment group.

Number of patients randomized and treated: 112 (28 patients are stillongoing)

Number of patients analyzed: 73 treated patients (37 Anti-EpCAM highdose, 36 Anti-EpCAM low dose)

Data Analyzed for the Foregoing Analysis:

All data leading to these results were monitored according toGCP-requirements but without data cleaning complete. The CRFs of allpatients completing all visits until week 12 and having all requiredtumor assessments at week 6 and 12 (n=23) were subject of a medicalreview with the emphasis to detect major protocol deviations and toclean the safety data. For these 23 patients the radiologic data werecentrally reviewed for this analysis.

Definition of Populations for Analysis:

-   -   Safety Analysis Set (SAF): All patients who received at least        one dose of the assigned study medication.    -   Full Analysis Set (FAS): Patients from the Safety Analysis Set        who had EpCAM positive (low/moderate or high expression) tumors        and at least one tumor assessment after start of therapy in case        of early withdrawal due to other reasons than clinical disease        progression.

The analysis of safety data was based on the SAF.

The analysis of baseline data and efficacy endpoints was based on thefull analysis sets.

Analyzed Endpoints (cf. Example 1.1, above):

-   -   Best Overall Tumor Response (OTR) rate at Week 12 (patients with        complete remission [CR] or partial remission [PR] according to        RECIST)    -   Clinical Benefit Rate (CBR) at Week 12 (patients with stable        disease or complete remission [CR] or partial remission [PR]        according to RECIST)    -   Clinical Benefit Rate (CBR) at Week 24 (patients with stable        disease or complete remission [CR] or partial remission [PR]        according to RECIST)    -   Time to Progression (TTP) as time from randomization and        alternatively time from start of treatment.        Subgroups Analyzed:

High, moderate and low EPCAM expressors were identified according toGastl et al. (2000). Lancet 356, 1981-2.

The following subgroups were analyzed:

-   -   EpCAM low/moderate expressors treated with low dose Anti-EpCAM        (“low dose Anti-EpCAM/low EpCAM”)    -   EpCAM low/moderate expressors treated with high dose Anti-EpCAM        (“high dose Anti-EpCAM/low EpCAM”)    -   EpCAM high expressors treated with low dose Anti-EpCAM (“low        dose Anti-EpCAM/high EpCAM”)    -   EpCAM high expressors treated with high dose Anti-EpCAM (“high        dose Anti-EpCAM/high EpCAM”)    -   EpCAM low/moderate expressors treated with low or high dose        Anti-EpCAM (“low EpCAM”)    -   EpCAM high expressors treated with low or high dose Anti-EpCAM        (“high EpCAM”)    -   Low dose Anti-EpCAM in EpCAM low/moderate or EpCAM high        expressors (“low dose Anti-EpCAM”)    -   High dose Anti-EpCAM in EpCAM low/moderate or EpCAM high        expressors (“high dose Anti-EpCAM”).        Analyses Performed:

Response (CBR or OTR) endpoints as defined above were analyzed asfollows:

-   -   CBR/OTR rate for each subgroup (see above) and all patients.    -   Fisher's exact test for CBR/OTR rate for the following        comparisons:    -   each subgroup (see above) vs. all other patients combined    -   “low dose Anti-EpCAM/low EpCAM” vs. “low dose Anti-EpCAM/high        EpCAM”    -   “low dose Anti-EpCAM/low EpCAM” vs. “high dose Anti-EpCAM/low        EpCAM”    -   “high dose Anti-EpCAM/high EpCAM” vs. “low dose Anti-EpCAM/high        EpCAM”    -   “high dose Anti-EpCAM/high EpCAM” vs. “high dose Anti-EpCAM/low        EpCAM”.

Time to clinical disease progression was defined as duration between thedate of the first Anti-EpCAM infusion (alternatively in a sensitivityanalysis: date of randomization) and the date of clinical diseaseprogression, i.e. the first incidence of progressive disease,respectively. If no clinical disease progression was observed, therespective time span was censored with the date of study termination. Incase of a missing date for study termination, the date of the last visitperformed was used instead.

Time-To-Progression (TTP) endpoints were analyzed as follows:

-   -   Median TTP (if estimable) for each subgroup and all patients.    -   Log-Rank test for TTP for the following comparisons:        -   each subgroup vs. all other patients combined        -   “low dose Anti-EpCAM/low EpCAM” vs. “low dose            Anti-EpCAM/high EpCAM”        -   “low dose Anti-EpCAM/low EpCAM” vs. “high dose            Anti-EpCAM/low EpCAM”        -   “high dose Anti-EpCAM/high EpCAM” vs. “low dose            Anti-EpCAM/high EpCAM”        -   “high dose Anti-EpCAM/high EpCAM” vs. “high dose            Anti-EpCAM/low EpCAM”.

Example 2.2 Results—Study Patients

Datasets Analyzed

Populations for analysis are tabulated in Table 4.

The full analysis set (FAS) (n=67) represents the analysis populationfor response and time to progression assessment. EpCAM status wasequally distributed in the two Anti-EpCAM dose groups, with about 38% oflow-EpCAM expressors and 57% of high EpCAM expressors in each treatmentgroup.

TABLE 4 Populations for Analysis¹ Treatment Group Low High Dose DoseAnti- Anti- Total EpCAM EpCAM (N = 73) (N = 36) (N = 37) Patients n % n% n % Total Safety Analysis Set (SAF) 73 100.0 36 100.0 37 100.0 FullAnalysis Set (FAS)² 67 91.8 32 88.9 35 94.6 EpCAM Negative Total 3 4.1 12.8 2 5.4 Safety Analysis Set (SAF) 3 100.0 1 100.0 2 100.0 FullAnalysis Set (FAS)² 0 0.0 0 0.0 0 0.0 EpCAM Low/Intermediate Total 2838.4 14 38.9 14 37.8 Safety Analysis Set (SAF) 28 100.0 14 100.0 14100.0 Full Analysis Set (FAS)² 27 96.4 13 92.9 14 100.0 EpCAM High Total42 57.5 21 58.3 21 56.8 Safety Analysis Set (SAF) 42 100.0 21 100.0 21100.0 Full Analysis Set (FAS)² 40 95.2 19 90.5 21 100.0 ¹Number ofpatients included in the administrative data review (%) ²Full AnalysisSet (FAS): Patients from the Safety Analysis Set who had EpCAM positive(low/moderate or high expression) tumors and at least one tumorassessment after start of therapy in case of early withdrawal due toother reasons than clinical disease progression.

Example 2.3 Results—Efficacy Analysis for Overall Tumor Response (OTR)and Clinical Benefit Rate (CBR)

According to the study protocol the primary endpoint of the study is theclinical benefit rate at week 24.

“Best Overall Tumor Response (OTR)”

The results for the “Best Overall Tumor Response (OTR) rate at Week 12”,the “Clinical Benefit Rate (CBR) at Week 12” and the “Clinical BenefitRate (CBR) at Week 24” in the FAS for EpCAM high and low expressors andoverall are presented in the following Table 5 and Table 6.

Response in terms of PR or CR according to RECIST criteria could not beconfirmed in any patient of the FAS in the central radiologicassessment.

“Clinical Benefit Rate (CBR)” at W12

Overall, 16 out of 67 patients (24%) of the FAS showed diseasestabilization (SD) at week 12. No significant difference betweentreatment groups was seen with regard to the clinical benefit rate atweek 12, which was 21.9% in the low dose group and 25.7% in the highdose group. The CBR was higher in the high EpCAM groups than in the lowEpCAM groups, however this difference was statistically not significant.

The Clinical Benefit Rate only comprised patients with stable disease asremissions according to RECIST could not be detected (see above).

TABLE 5 Clinical Benefit Rate (CBR) at Week 12 (patients with stabledisease or complete remission [CR] or partial remission [PR] accordingto RECIST) - Full Analysis Set Low High Dose Anti- Dose Anti- EpCAM¹EpCAM¹ n % n % P-Value² Low EpCAM (N³ = 13/14) 2 15.4 2 14.3 1.0 HighEpCAM (N³ = 19/21) 5 26.3 7 33.3 0.7365 Overall (N³ = 32/35) 7 21.9 925.7 0.7797 ¹Number of patients with clinical benefit among patients forwhich clinical benefit could be assessed according to RECIST ²Two-sidedFisher's Exact Test ³N in low dose group/high dose group“Clinical Benefit Rate (CBR)” at W24

A higher clinical benefit rate was seen at week 24 in the high dosegroup (14.3%) as compared to the low dose group (6.3%). In the high dosegroup the CBR is identical (14.3%) for the low and high EpCAM subgroups.In the low dose group the CBR is similar for the high and low EpCAMexpressors (5.3 vs. 7.7%).

The CBR is only based on patients with stable disease as no PR or CRwere detected.

TABLE 6 Clinical Benefit Rate (CBR) at Week 24 (patients with stabledisease or complete remission [CR] or partial remission [PR] accordingto RECIST) Low High Dose Anti- Dose Anti- EpCAM¹ EpCAM¹ n % n % P-Value²Low EpCAM (N³ = 13/14) 1 7.7 2 14.3 1.0 High EpCAM (N³ = 19/21) 1 5.3 314.3 0.6039 Overall (N³ = 32/35) 2 6.3 5 14.3 0.4266 ¹Number of patientswith clinical benefit among patients for which clinical benefit could beassessed according to RECIST ²Two-sided Fisher's Exact Test ³N in lowdose group/high dose groupEfficacy Analysis: Time to Progression

Time to clinical disease progression was defined as duration between thedate of the first Anti-EpCAM infusion (alternatively in the sensitivityanalysis: date of randomization) and the date of clinical diseaseprogression, i.e. the first incidence of progressive disease accordingto RECIST. If no clinical disease progression was observed, the timespan to the date of study termination was taken. In case the date ofstudy termination was not available, the date of the last visitperformed was taken instead.

Median time to progression in the FAS for the low and high dose groupand the EpCAM subgroups is presented in Table 7 (time from start oftreatment to clinical disease progression) and Table 8 (time fromrandomization to clinical disease progression).

Overall Anti-EpCAM high dose showed a clear prolongation of the mediantime to progression (calculated as time from first infusion) from 43 to78 days when compared to the Anti-EpCAM low dose treatment (as shown inFIG. 2). This difference was statistically significant when testing the“survival” curves (p=0.0348; log rank test) (as shown in FIG. 3).Similar, a difference in median time to progression (calculated as timefrom first infusion) was observed when comparing patients with lowversus high EpCAM expression (42 to 80 days, respectively; p=0.0431; logrank test). The highest median time to progression (calculated as timefrom first infusion) was observed in patients with high EpCAM expressiontreated with high dose of Anti-EpCAM (90 days; p=0.0238; log ranktest—compared to all other patients) (as shown in FIG. 5).

TABLE 7 Median Time to Progression (time from start of infusion toclinical disease progression [days]) - Full Analysis Set Low Dose HighDose Anti-EpCAM¹ Anti-EpCAM¹ P-Value² Low EpCAM 41 days 47 days 0.1451(N³ = 13/14) High EpCAM 49 days 90 days 0.1262 (N³ = 19/21) Overall (N³= 32/35) 43 days 78 days 0.0348 ¹Median time to progression in days ²LogRank Test on differences between treatment groups ³N in low dosegroup/high dose group

The data of Table 7 are graphically represented in FIG. 4.

For the time to progression calculated from the day of randomizationcomparable results were found. Again overall Anti-EpCAM high dose showeda clear prolongation of the median time to progression (calculated astime from randomization) from 46 to 79 days in comparison to theAnti-EpCAM low dose treatment. This difference was statisticallysignificant when testing the “survival” curves (p=0.0441; log ranktest). The treatment difference was also more pronounced in the highEpCAM group with median times to progression of 63 and 91 days for thelow dose and high dose high EpCAM groups, respectively, and 43 and 53days in the low dose and high dose low EpCAM group, respectively.

TABLE 8 Median Time to Progression (time from randomization to clinicaldisease progression [days]) - Full Analysis Set Low Dose High DoseAnti-EpCAM¹ Anti-EpCAM¹ P-Value² Low EpCAM (N³ = 13/14) 43 days 53 days0.2353 High EpCAM (N³ = 19/21) 63 days 91 days 0.1350 Overall (N³ =32/35) 46 days 79 days 0.0441Efficacy Conclusions

Based on the study protocol

-   -   The best overall response (OTR)    -   The clinical benefit rate (CBR); and    -   The time to progression (TTP)        were analyzed comparing the high dose Anti-EpCAM group and the        low dose Anti-EpCAM group in the overall population and he low-        and high-EpCAM subgroups.

The clinical benefit rates at weeks 12 and 24 (W12 and W24,respectively) could be established comprising all patients showingstable disease at the respective time points.

-   -   At W12 the CBR was slightly higher in the high dose Anti-EpCAM        group than in the low dose group (25.7% vs. 21.9%), also showing        higher rates in both dosage groups for the high EpCAM subgroups.    -   At W24 the CBR was higher in the high dose Anti-EpCAM group        (14.3% vs. 6.3% in the low dose Anti-EpCAM group). No marked        differences between the EpCAM subgroups could be detected.

The median time to progression in the overall sample showed a clearprolongation for the Anti-EpCAM high dose compared to the Anti-EpCAM lowdose (43 to 78 days), the difference being statistically significantwhen testing the survival curves (p=0.0348, log rank test). The mediantime to progression (calculated as time from first infusion) wasobserved in patients with high EpCAM expression treated with high doseof Anti-EpCAM (90 days; p=0.0238; log rank test—compared to all otherpatients).

Overall Conclusions

The data available showed long-term disease stabilization (>week 24) inat least 7 patients with some patients still ongoing.

As is clearly visible in FIGS. 2-5, the time to progression evaluationshowed a pronounced prolongation of “survival time” in favor of the highdose Anti-EpCAM population reaching statistical significance.Specifically, as shown in FIG. 5, the patient with high EpCAM expressionreceiving high doses of Anti-EpCAM significantly prolongedprogression-free survival (90 days versus 41-49 days in the othergroups).

Example 3 Final Study Report for Phase II Clinical study of Anti-EpCAMExample 3.1 Summary and Introduction to Study

The clinical study was performed as described above in Example 1. Theresults of this study follow now in Example 3.

Methodology:

A randomized, open-label, multicenter, parallel group, phase II study.The study was designed to evaluate the efficacy and safety of Anti-EpCAMover 24 weeks of therapy at two different doses with positive EpCAMtesting. The central randomization process was stratified according tothe EpCAM test results performed at screening. Upon registration in oneof the EpCAM strata, patients were randomly assigned to either the lowdose treatment group or the high dose treatment group.

Number of patients randomized and treated: 112

Number of patients analyzed: 112 treated patients (56 Anti-EpCAM highdose, 56 Anti-EpCAM low dose), thereof 109 patients tested EpCAM+.

Data analyzed for the Foregoing Analysis:

All data leading to these results were monitored and cleaned accordingto GCP-requirements and the database was locked before the finalanalysis was performed.

Definition of Populations for Analysis:

-   -   Safety Analysis Set (SAF): All patients who received at least        one dose of the assigned study medication.    -   Full Analysis Set (FAS): Patients from the Safety Analysis Set        who had EpCAM positive (low/moderate or high expression) tumors        and at least one tumor assessment after start of therapy in case        of early withdrawal due to other reasons than clinical disease        progression.

The analysis of safety data was based on the SAF.

The analysis of baseline data and efficacy endpoints was based on thefull analysis set.

Analyzed Endpoints (cf. Example 1.1, above):

-   -   Clinical Benefit Rate (CBR) at Week 24 (patients with stable        disease [SD] or complete remission [CR] or partial remission        [PR] according to RECIST)    -   Best Overall Tumor Response (OTR) rate at Week 12 (patients with        complete remission [CR] or partial remission [PR] according to        RECIST)    -   Clinical Benefit Rate (CBR) at Week 12 (patients with stable        disease [SD] or complete remission [CR] or partial remission        [PR] according to RECIST)    -   Time to Progression (TTP) as time from randomization and        alternatively time from start of treatment.        Subgroups Analyzed:

High and low/moderate EPCAM expressors were identified according toGastl et al. (2000). Lancet 356, 1981-2.

The following subgroups were analyzed:

-   -   EpCAM low/moderate expressors treated with low dose Anti-EpCAM        (“low dose Anti-EpCAM/low EpCAM”)    -   EpCAM low/moderate expressors treated with high dose Anti-EpCAM        (“high dose Anti-EpCAM/low EpCAM”)    -   EpCAM high expressors treated with low dose Anti-EpCAM (“low        dose Anti-EpCAM/high EpCAM”)    -   EpCAM high expressors treated with high dose Anti-EpCAM (“high        dose Anti-EpCAM/high EpCAM”)    -   EpCAM low/moderate expressors treated with low or high dose        Anti-EpCAM (“low EpCAM”)    -   EpCAM high expressors treated with low or high dose Anti-EpCAM        (“high EpCAM”)    -   Low dose Anti-EpCAM in EpCAM low/moderate or EpCAM high        expressors (“low dose Anti-EpCAM”)    -   High dose Anti-EpCAM in EpCAM low/moderate or EpCAM high        expressors (“high dose Anti-EpCAM”).        Analyses Performed:

Response (CBR or OTR) endpoints as defined above were analyzed asfollows:

-   -   CBR/OTR rate for each subgroup (see above) and all patients.

Time to clinical disease progression was defined as duration between thedate of the first Anti-EpCAM infusion (alternatively in a sensitivityanalysis: date of randomization) and the date of clinical diseaseprogression, i.e. the first incidence of progressive disease,respectively. If no clinical disease progression was observed, therespective time span was censored with the date of study termination. Incase of a missing date for study termination, the date of the last visitperformed was used instead.

Time-To-Progression (TTP) endpoints were analyzed as follows:

-   -   Median TTP (if estimable) for each subgroup and all patients.    -   Log-Rank test for TTP on differences between the treatment        groups for the following comparisons:        -   each subgroup vs. all other patients combined        -   “low dose Anti-EpCAM/low/moderate EpCAM” vs. “low dose            Anti-EpCAM/high EpCAM”        -   “low dose Anti-EpCAM/low/moderate EpCAM” vs. “high dose            Anti-EpCAM/low/moderate EpCAM”        -   “high dose Anti-EpCAM/high EpCAM” vs. “low dose            Anti-EpCAM/high EpCAM”        -   “high dose Anti-EpCAM/high EpCAM” vs. “high dose            Anti-EpCAM/low/moderate EpCAM”.        -   “low dose Anti-EpCAM/low/moderate EpCAM” vs. “high dose            Anti-EpCAM/high EpCAM”        -   “low dose Anti-EpCAM” vs. “high dose Anti-EpCAM”        -   “high EpCAM” vs. “low/moderate EpCAM”

Example 3.2 Results—Study Patients

Datasets Analyzed

Populations for analysis are tabulated in Table 9.

The full analysis set (FAS) (n=109) represents the analysis populationfor response and time to progression assessment.

TABLE 9 Populations for Analysis 1 Low Dose MT201 High Dose MT201 (N =55) (N = 54) Low/Moderate High Low/Moderate High EpCAM (N = 19) EpCAM (N= 36) EpCAM (N = 16) EpCAM (N = 38) n % n % n % n % Race² Caucasian 19100 36 100 16 100 38 100 Age [years]¹ n = 19 n = 36 n = 16 n = 38 60.3 ±9.8 59.2 ± 9.6 57.8 ± 11.5 59.1 ± 11.5 (42.0-79.0, 61.0) (42.0-75.0,61.5) (40.0-80.0, 54.5) (38.0-81.0, 60.5) Height [cm]¹ n = 19 n = 36 n =16 n = 38 160.8 ± 7.9 161.8 ± 6.6 160.4 ± 6.9 162.4 ± 7.5 (149.0-177.0,161.0) (150.0-175.0, 162.5) (150.0-170.0, 161.5) (145.0-176.0, 162.5)Weight [kg]¹ n = 19 n = 36 n = 16 n = 38 67.1 ± 6.5 70.1 ± 13.2 68.1 ±7.9 69.0 ± 15.3 (57.0-75.0, 69.0) (50.0-108.0, 68.5) (53.5-84.0, 66.5)(42.7-115.0, 65.0) BMI [kg/m²]¹ n = 19 n = 36 n = 16 n = 38 26.1 ± 3.126.8 ± 4.8 26.7 ± 4.1 26.2 ± 5.9 (20.4-31.6, 26.2) (19.3-39.7, 26.7)(18.7-33.8, 27.2) (17.1-49.1, 25.4) ¹Number of patients, mean ± standarddeviation (minimum-maximum, median) ²Number of patients, %

Example 3.3 Results—Efficacy Analysis for Overall Tumor Response (OTR)and Clinical Benefit Rate (CBR)

According to the study protocol the primary endpoint of the study is theclinical benefit rate at week 24.

“Best Overall Tumor Response (OTR)”

The results for the “Best Overall Tumor Response (OTR) rate at Week 12”,the “Clinical Benefit Rate (CBR) at Week 12” and the “Clinical BenefitRate (CBR) at Week 24” in the FAS for EpCAM high and low expressors andoverall are presented in the following Table 10 and Table 11.

Two responses (in terms of PR or CR according to RECIST criteria) werediagnosed by local radiologic assessments but could not be confirmed inany patient of the FAS in the central radiologic assessment.

“Clinical Benefit Rate (CBR)” at W24

A trend towards higher clinical benefit rate (CBR), albeit notsignificant, was seen at week 24 in the high dose group (7.9%) ascompared to the low dose group (4.5%). Similarly, the CBR for the highEpCAM expressors shows a trend towards higher CBR rates as compared tolow/moderate EpCAM expressors (7.3% vs. 3.7%).

The CBR is only based on patients with stable disease as no PR or CRwere detected.

TABLE 11 Clinical Benefit Rate (CBR) at Week 24 (patients with stabledisease or complete remission [CR] or partial remission [PR] accordingto RECIST) Low Dose MT201 High Dose MT201 (N = 44) (N = 38) Low/ModerateHigh Low/Moderate High EpCAM (N = 15) EpCAM (N = 29) EpCAM (N = 12)EpCAM (N = 26) n¹ %¹ n¹ %¹ n¹ %¹ n¹ %¹ Clinical Benefit² (CR, PR and SD)0 0.0 2 6.9 1 8.3 2 7.7 No Clinical Benefit² (PD and Not Available) 15100.0 26 89.7 11 91.7 24 92.3 Not Evaluable^(2,3) 0 0.0 1 3.4 0 0.0 00.0 Lower 95% Confidence Limit for CBR 0.0% 1.2% 0.4% 1.4% ¹Number ofpatients (%) ²According to ‘final’ assessment ³patients still in thestudy with respective response assessment classified as ‘not assessable’in the central/IRAB assessment“Clinical Benefit Rate (CBR)” at W12

Overall, 17 out of 109 patients (16%) of the FAS showed diseasestabilization (SD) at week 12. A trend towards higher clinical benefitrate (CBR), albeit not significant, was seen at week 12 in the high dosegroup (16.7%) as compared to the low dose group (14.5%). Similarly, theCBR for the high EpCAM expressors shows a trend towards higher CBR ratesas compared to low/moderate EpCAM expressors (18.9% vs. 8.6%).

TABLE 10 Clinical Benefit Rate (CBR) at Week 12 (patients with stabledisease or complete remission [CR] or partial remission [PR] accordingto RECIST) - Full Analysis Set Low Dose MT201 High Dose MT201 (N = 55)(N = 54) Low/Moderate High Low/Moderate High EpCAM (N = 19) EpCAM (N =36) EpCAM (N = 16) EpCAM (N = 38) n¹ %¹ n¹ %¹ n¹ %¹ n¹ %¹ ClinicalBenefit² (CR, PR and SD) 1 5.3 7 19.4 2 12.5 7 18.4 No Clinical Benefit²(PD and Not Available) 18 94.7 28 77.8 14 87.5 28 73.7 NotEvaluable^(2,3) 0 0.0 1 2.8 0 0.0 3 7.9 ¹Number of patients (%)²According to ‘final’ assessment ³patients still in the study withrespective response assessment classified as ‘not assessable’ in thecentral/IRAB assessmentEfficacy Analysis: Time to Progression

Time to clinical disease progression was defined as duration between thedate of the first Anti-EpCAM infusion (alternatively in the sensitivityanalysis: date of randomization) and the date of clinical diseaseprogression, i.e. the first incidence of progressive disease accordingto RECIST. If no clinical disease progression was observed, the timespan to the date of study termination was taken. In case the date ofstudy termination was not available, the date of the last visitperformed was taken instead.

Analysis of Kaplan-Maier (KM-) curves for time to progression ispresented in FIGS. 6-8 (time from start of treatment to clinical diseaseprogression).

Overall Anti-EpCAM high dose showed a significant prolongation of thetime to progression over time when compared to the Anti-EpCAM low dosetreatment (as shown in FIG. 6; Hazard ratio (HR)=0.666). This differencewas statistically significant when testing the “survival” curves(p=0.0465; log rank test). Similar, a trend towards a difference in timeto progression (calculated as time from first infusion) was observedwhen comparing patients with low/intermediate versus high EpCAMexpression (FIG. 7; HR=0.706; p=0.1157; log rank test). The highest riskreduction time to progression (calculated as time from first infusion)was observed in patients with high EpCAM expression treated with highdose of Anti-EpCAM (HR=0.433; p=0.0057; log rank test—compared topatients with low EpCAM expression treated with low dose of Anti-EpCAM)(as shown in FIG. 8).

Efficacy Conclusions

Based on the study protocol

-   -   The best overall response (OTR)    -   The clinical benefit rate (CBR); and    -   The time to progression (TTP)        were analyzed comparing the high dose Anti-EpCAM group and the        low dose Anti-EpCAM group in the overall population and the low-        and high-EpCAM subgroups.

The clinical benefit rates at weeks 12 and 24 (W12 and W24,respectively) could be established comprising all patients showingstable disease at the respective time points.

-   -   At W12 the CBR was slightly higher in the high dose Anti-EpCAM        group than in the low dose group (16.7% vs. 14.5%), also showing        higher rates in both dosage groups for the high EpCAM subgroups        (18.9% vs. 8.6% in the low/intermediate EpCAM expressors).    -   At W24 the CBR was higher in the high dose Anti-EpCAM group        (7.9% vs. 4.5% in the low dose Anti-EpCAM group) also indicating        higher rates in both dosage groups for the high EpCAM subgroups        (7.3% vs. 3.7% in the low/intermediate EpCAM expressors).

The time to progression analysis in the overall sample showed aprolongation for the Anti-EpCAM high dose compared to the Anti-EpCAM lowdose (R=0.666), the difference being statistically significant whentesting the survival curves (p=0.0465, log rank test). The highestprolongation time to progression (calculated as time from firstinfusion) was observed in patients with high EpCAM expression treatedwith high dose of Anti-EpCAM (HR=0.433; p=0.0057; log rank test—comparedto patients with low EpCAM expression treated with low dose ofAnti-EpCAM).

Overall Conclusions

The data available showed long-term disease stabilization (>week 24) inat least 6 patients according to central radiologic review.

As is clearly visible in FIGS. 6-8, the time to progression evaluationshowed a prolongation of “survival time” in favor of the high doseAnti-EpCAM population reaching statistical significance. Specifically,as shown in FIG. 8, patients with high EpCAM expression receiving highdoses of Anti-EpCAM significantly prolonged progression-free survival.

The invention claimed is:
 1. A method of treating human metastatic breast cancer in a subject said method comprising administering to a human an anti-EpCAM antibody comprising SEQ ID Nos. 3, 4, 5, 6, 7 and 8, wherein said subject exhibits high EpCAM expression showing a total immunostaining score for EpCAM of greater than 4, wherein said antibody is administered as at least one loading dose of 2 mg/kg body weight followed by multiple maintenance doses, each maintenance dose being 2 mg/kg body weight, wherein each loading dose is administered every week and each maintenance dose is administered every second week, wherein one loading dose is administered at the beginning of each of therapy weeks 1, 2 and 3 followed by 11 maintenance doses, and one maintenance dose is administered at the beginning of each of therapy weeks 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and
 24. 2. The method of claim 1, wherein the anti-EpCAM antibody comprises SEQ ID Nos. 1 and
 2. 3. The method of claim 1, wherein the treatment comprises long-term stabilization of metastatic breast cancer.
 4. The method according to claim 1, wherein metastatic breast cancer is classified as Stage IV according to the Tumor Node Metastasis (“TNM”) system.
 5. The method of claim 1, wherein the antibody is administered in a solution comprising 0.9% sodium chloride solution.
 6. The method of claim 1, wherein the antibody is administered intravenously. 